In article <kallisti-1707952017580001 at lomasneylab.feinberg.nwu.edu>,
kallisti at merle.acns.nwu.edu (Patrick Grealish) wrote:
> HEllo, I am current starting another experiment where i'll need to
produce
>eucarotic protein in E. coli.
>snip<
Hello Patrick,
Our lab uses the GST system from Pharmacia. You get a fusion protein with
Glutathione-S-Transferase and can cleave with your choice of proteases. I
have used pGEX-2T with success, although I have had problems getting my
protein cleaved away from the GST. I didn't need my protein absolutely
pure, so it wasn't important. Another person in the lab has used pGEX-2T &
pGEX-2TK with good success. The latter vector is usefull in that it has a
protein kinase site after the protease site, but in front of your protein
that you can lable stinking hot with gamma 32 ATP.
Good Luck
Brad
===========================================================================
Brad Nicholson |"If it worked the first time, it wouldn't be
Department of Pathology | research."...Brad Nicholson
University of Utah | Live from behind the Zion Curtian.
Salt Lake City, UT 84132 |
brad at corona.med.utah.edu |
or: (801)-581-4365 | My opinions are solely my own.
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